A prokaryotic expression vector for the mutant diphtheria toxin CRM197 was constructed and expressed in Esch-erichia coli cells.Anti-CRM197 antibody concentrations were detected in serum samples of healthy volunteers.The crm 197 gene was codon-optimized in E.coli and cloned into the plasmid pET28a(+)under optimized expression conditions.CRM197 was purified using Ni-NTA spin columns and ion exchange chromatography,and confirmed by western blot analysis.The puri-fied CRM197 was used to detect specific anti-CRM197 antibody levels in serum samples of different age groups.The results showed that soluble codon-optimized CRM197 was successfully expressed under optimized expression conditions.The purity of CRM197 was more than 95％,as determined with Ni-NTA spin columns and ion exchange chromatography,consistent with the single specific bands obtained by western blot analysis and detection of serum levels of the anti-CRM197 antibody.Collec-tively,these results confirmed that the proposed expression strategy achieved high-yield production of soluble CRM197,al-though high levels in human serum may affect evaluation of immune interactions with glycan-CRM197 conjugates for applica-tion as a diagnostic antigen.The diphtheria mutant toxin CRM197 is used in many conjugate vaccines.The synthetic crm 197 gene with codon optimization in pET28a was transformed into E.coli Origami B(DE3)cells.CRM197 was induced by isopro-pyl β-d-1-thiogalactopyranoside and high level accumulation of soluble CRM197 was purified using Ni-NTA spin columns and ion exchange chromatography.The purity of the final prepara-tion reached 95％.CRM197 was used to detect the concentra-tions of the anti-CRM197 antibody in serum samples of healthy volunteers of different ages.The proposed expression strategy yielded high production of CRM197,which could interfere with evaluations of induced immune interactions by glycan-CRM197 conjugates and prohibit application as a diagnostic antigen.

Objective:To understand the serological characteristics of irregular antibodies in pregnant women and explore their clinical significance.Methods:From January 2017 to March 2022,151 471 pregnant women in Women and Children's Hospital of Chongqing Medical University were enrolled in this study,microcolumn gel card test was used for irregular antibody screening,and antibody specificity identification was further performed in some antibody-positive subjects.Results:The positive rate of irregular antibody screening in the enrolled pregnant women was 0.91％(1 375/151 471),0.23％(355/151 471)was detected in the first trimester,0.05％(71/151 471)in the second trimester,and 0.63％(949/151 471)in the third trimester.The positive rate of irregular antibody screening in the third trimester was significantly higher than that in the first and second trimester,and a significant increase in the number of positive cases was found in the third trimester than that in the second trimester.The analysis of agglutination intensity of 1 375 irregular antibody screening positive results showed that the weakly positive agglutination intensity accounted for 50.11％(689/1 375),which was the highest,the suspicious positive was 18.69％(257/1 375),and the positive was 31.20％(429/1 375).The significant difference in distribution of agglutination intensity was not observed between the first trimester group and the second trimester group,however,in the third trimester,the proportion of suspicious positive and weakly positive was lower than the first trimester,while,the proportion of positive was higher than the first trimester,and the difference was statistically significant(P＜0.001).Among the irregular antibody screening positive pregnant women,the proportion of pregnant women with pregnancy number ≥ 2 was significantly higher than that with pregnancy≤1.Among 60 pregnant women who underwent antibody identification,the distributions of the antibodies were as follows:Rh blood group system accounted for 23.33％(14/60),Lewis system 43.33％(26/60),Kidd system 3.33％(2/60),MNS system 16.67％(10/60),P1PK system 1.67％(1/60),autoantibodies 1.67％(1/60),and 4 cases was unable to identify(6.67％,4/60).Among specific antibodies,the anti-Lea was the most common(30.00％),followed by anti-E(16.67％)and anti-M(16.67％).Conclusion:The differences of irregular antibody serological characteristics exist in pregnant women from different regions with different genetic backgrounds,understanding the characteristics of irregular antibody in local pregnant women is of great significance for ensuring transfusion safety in pregnant women and preventing hemolytic disease of newborn.

Objective To develop a portable field infusion kit for first aid to meet the treatment requirements in field conditions.Methods The infusion kit had its frame made of third-generation carbon fiber material and was composed of a lid and a body.The lid was equipped with embedded handles on the outer surface,a medicine module consisting of 12 mesh bags on the inner surface and LED lights on the inner and outer front side walls.The kit body had three storage layers in its upper,middle and lower parts,of which the upper layer was for supplies of infusion and hemostatic dressing,the middle layer was for medical instruments and soft bags of liquid for first aid,and the lower layer was for medical waste and sharp instruments,and the bottom of the kit was provided with telescopic drawbars and invisible rollers.Results The first-aid infusion kit facilitated the storage and utilization of kinds of infusion supplies,and lowered time consumption and workloads for transport.Conclusion The first-aid infusion kit gains advantages in reasonable layout,comprehensive functions and convenient operation,and thus is worhty promoting for casualty treatment in field conditions.[Chinese Medical Equipment Journal,2024,45(7):115-117]

Objective To study the clinical manifestations,laboratory features,and labial gland pathological features in children with systemic lupus erythematosus(SLE)complicated by Sj?gren's syndrome(SS).Methods A retrospective analysis was conducted on 102 children with SLE who underwent labial gland biopsies at Renji Hospital,Shanghai Jiao Tong University School of Medicine from January 2013 to December 2022.The children were divided into two groups based on the presence of SS:the SLE with SS group(SLE-SS;60 children)and the SLE-only group(42 children).According to the focus score(FS)of the labial glands,children in the SLE-SS group were further subdivided into FS≥4 subgroup(26 children)and FS<4 subgroup(34 children).The clinical data of the groups were compared.Results Compared to the SLE-only group,children in the SLE-SS group had less skin and mucosal involvement,were more likely to have positive anti-SSA and anti-SSB antibodies,and had higher levels of rheumatoid factor(P<0.05).There was no significant difference in treatment protocols between the two groups(P>0.05).Compared to the FS<4 subgroup,the FS≥4 subgroup had more frequent musculoskeletal involvement(P<0.05),but there was no significant difference in SLE disease activity or other major organ involvement between the subgroups(P>0.05).Conclusions Children with SLE complicated by SS are less likely to have skin and mucous membrane involvement and exhibit specific serological characteristics.The SLE-SS children with an FS≥4 are more likely to experience musculoskeletal involvement.However,FS is not associated with disease activity or other significant organ damage.

This article reports on the clinical and genetic characteristics of monozygotic twins with Marshall-Smith syndrome(MRSHSS)due to a mutation in the NFIX gene,along with a review of related literature.Both patients presented with global developmental delays,a prominent forehead,shallow eye sockets,and pectus excavatum.Genetic testing revealed a heterozygous splicing site mutation c.697+1G>A in both children,with parents showing wild-type at this locus.According to the guidelines of the American College of Medical Genetics and Genomics,this mutation is considered likely pathogenic and has not been previously reported in the literature.A review of the literature identified 32 MRSHSS patients with splicing/frameshift mutations.Accelerated bone maturation and moderate to severe global developmental delay/intellectual disability are the primary clinical manifestations of patients with MRSHSS.Genetic testing results are crucial for the diagnosis of this condition.

A boy,aged 7 months,presented with severe global developmental delay(GDD),refractory epilepsy,hypotonia,nystagmus,ocular hypertelorism,a broad nasal bridge,everted upper lip,a high palatal arch,and cryptorchidism.Genetic testing revealed a de novo heterozygous missense mutation of c.364G>A(p.E122K)in the EEF1A2 gene,and finally the boy was diagnosed with autosomal dominant developmental and epileptic encephalopathy 33 caused by the EEF1A2 gene mutation.This case report suggests that for children with unexplained infancy-onset severe to profound GDD/intellectual disability and refractory epilepsy,genetic testing for EEF1A2 gene mutations should be considered.This is particularly important for those exhibiting hypotonia,nonverbal communication,and craniofacial deformities,to facilitate a confirmed diagnosis.

Aim To investigate the mechanism by which formononetin (FN) inhibits mitochondrial dynamic-related protein 1 (DRP1) -NLRP3 axis via intervening the generation of ROS to reduce allergic airway inflammation. Methods In order to establish allergic asthma mouse model, 50 BALB/c mice aged 8 weeks were divided into the control group, model group, FN treatment group and dexamethasone group after ovalbumin (OVA) induction. Airway inflammation and collagen deposition were detected by HampE and Masson staining. Th2 cytokines and superoxide dismutase (SOD), catalase (CAT), malondialdehyde (MDA), and IgE levels in bronchoalveolar lavage fluid (BALF) were measured by ELISA, ROS in BEAS-2B cells was assessed by DCFH-DA staining, DRP1 expression in lung tissue and BEAS-2B cells was detected by immunohistochemistry and immunofluorescence, and the DRP1-NLRP3 pathway was analyzed by immunoblotting. Results FN treatment could effectively ameliorate the symptoms of asthmatic mouse model, including reducing eosinophil accumulation, airway collagen deposition, decreasing Th2 cytokine and IgE levels, reducing ROS and MDA production, increasing SOD and CAT activities, and regulating DRP1-NLRP3 pathway-related protein expression, thereby relieving inflammation. Conclusion FN ameliorates airway inflammation in asthma by regulating DRP1-NLRP3 pathway.

AIM: To explore the protective effect of astragalus glycyrrhiza decoction (AGD) on arsenic trioxide (ATO)-induced QT interval prolongation and its mechanism based on metabonomics. METHODS: The model of ATO-induced QT interval prolongation in rats was established, and ECG, blood routine, and metabonomics were detected, and the key targets were collected combined with network pharmacology. The possible candidate genes and pathways for the protective effect of AGD were screened by GO and KEGG enrichment analysis and then verified by experiments in vitro. RESULTS: AGD could significantly alleviate the ATO-induced QT interval of SD rats. GO enrichment analysis was mainly related to inflammatory response, reactive oxygen species, oxidative stress, inner cell vesicles, folds, inner cell vesicles, SMAD binding, R-SMAD binding, and signal receptor activator activity. KEGG analysis showed that it was mainly concentrated in the PI3K-Akt signal pathway, lipid and arteriosclerosis, FOXO signal pathway, TNF signal pathway, HIF-1, and other signal pathways. Through the H9c2 cell model in vitro, it was verified that AGD could reverse the expression of SIRT1 and FOXO1 proteins. CONCLUSION: AGD may improve the ATO-induced QT interval prolongation and reduce the cardiotoxicity of ATO by regulating the SIRT1 / FOXO1 signal pathway.

Objective To observe the clinical efficacy of Qigui Tongluo Oral Liquid(a hospital preparation developed by Guangdong Second Traditional Chinese Medicine Hospital and being composed of Astragali Radix,Angelicae Sinensis Radix,Notoginseng Radix et Rhizoma,Paeoniae Radix Rubra,Achyranthis Bidentatae Radix,Spatholobi Caulis,Salviae Miltiorrhizae Radix et Rhizoma,Pheretima,etc.)combined with moxibustion for the treatment of chronic fatigue syndrome(CFS)of qi deficiency and blood stasis type on the basis of qi-collateral theory.Methods A retrospective study was conducted in 60 CFS patients with qi deficiency and blood stasis type.The patients were divided into an observation group and a control group,with 30 patients in each group according to the therapy.The control group was treated with mild moxibustion on Shenque(CV8)point with moxa sticks,and the observation group was treated with Qigui Tongluo Oral Liquid on the basis of treatment for the control group.The course of treatment lasted for 4 weeks.The changes of traditional Chinese medicine(TCM)syndrome scores,Fatigue Scale-14(FS-14)scores,serum immunoglobulin IgA,IgM,IgG levels,and cortisol(COR)level in the two groups were observed before and after the treatment.After treatment,the clinical efficacy and safety of the two groups were evaluated.Results(1)After 4 weeks of treatment,the total effective rate of the observation group was 96.67%(29/30),and that of the control group was 80.00%(24/30).The intergroup comparison showed that the clinical efficacy of the observation group was significantly superior to that of the control group(P＜0.05).(2)After treatment,the TCM syndrome scores and FS-14 scores of patients in the two groups were significantly decreased compared with those before treatment(P＜0.01),and the effect on decreasing TCM syndrome scores and FS-14 scores in the observation group was significantly superior to that in the control group(P＜0.01).(3)After treatment,the serum IgA and IgG levels of the two groups as well as the serum IgM and COR levels of the observation group were significantly increased compared with those before treatment(P＜0.01),and the effect of the observation group on increasing serum IgA,IgM,IgG,and COR levels was significantly superior to that of the control group(P＜0.05 or P＜0.01).(4)During the treatment,there were no significant adverse reactions occurring in the two groups.Conclusion Healthy-qi deficiency and collateral obstruction contribute to the core pathogenesis of CFS.Based on the TCM qi-collateral theory and following the therapeutic principle of replenishing deficiency and unblocking collaterals,Qigui Tongluo Oral Liquid combined with moxibustion for the treatment of CFS patients with qi deficiency and blood stasis type can achieve certain efficacy.The combined therapy could significantly alleviate the clinical symptoms,improve the immunity level,and regulate the neuro-endocrine-immune(NEI)network of the patients.

Objective:To establish an ultrasound prediction model of postoperative recurrence in patients with papillary thyroid carcinoma(PTC)after complete endoscopic radical surgery.Meth-ods:264 patients with PTC who underwent complete endoscopic radical surgery for the first time in our hospital from February 2017 to March 2020 were retrospectively collected.They were divided in-to recurrence group and non recurrence group according to whether there was recurrence after surgery.The clinical data,nodule diameter,nodule number,internal echo,microcalcification and other ultrasonic data of the two groups were compared.Single factor,Lasso and Logistic regression mod-els were used to analyze the influencing factors of postoperative recurrence of PTC patients,and an nomogram model was established based on the selected indicators.Results:Compared with the non recurrence group,the patients in the recurrence group had larger nodule diameter,irregular nod-ule edge,aspect ratio＞1,microcalcification and capsule invasion(P＜0.05).Nodular diameter＞10 mm,irregular edge,aspect ratio＞1,microcalcification and capsule invasion were independent risk factors for postoperative recurrence of PTC patients(P＜0.05).The C-index of the constructed nomogram model was 0.756(95％Cl:0.684～0.830),and the AUC of the ROC curve was 0.895(95％Cl:0.866～0.915);The calibration curve results show that the average deviation is 0.027,and the predic-tion probability fits the actual probability well;The clinical decision curve is far away from the extreme curve and has good clinical applicability.Conclusion:The nomogram model based on nodule size,irregular margin,microcalcification,aspect ratio＞1,and capsule invasion has good accuracy in pre-dicting the recurrence of PTC patients after complete endoscopic radical surgery,and has certain clinical significance.